Lipase inhibiting polymers

ABSTRACT

The invention features a method for treating obesity in a patient by administering to the patient a polymer that has been substituted with one or more groups that inhibit lipases, which are enzymes responsible for the hydrolysis of fat. The invention further relates to the polymers employed in the methods described herein as well as novel intermediates and methods for preparing the polymers.

RELATED APPLICATION

[0001] This application is a Divisional of U.S. application Ser. No.09/714,541, filed Nov. 16, 2000, which is Divisional of U.S. applicationSer. No. 09/226,585, filed Jan. 6, 1999, now U.S. Pat. No. 6,352,692,which is a Continuation-in-Part of U.S. application Ser. No. 09/166,510filed Oct. 5, 1998, now U.S. Pat. No. 6,267,952, which is aContinuation-in-Part of U.S. application Ser. No. 09/005,379 filed onJan. 9, 1998, now abandoned, the entire teachings of which areincorporated herein by reference.

BACKGROUND OF THE INVENTION

[0002] Human obesity is a recognized health problem with approximatelyninety-seven million people considered clinically overweight in theUnited States. The accumulation or maintenance of body fat bears adirect relationship to caloric intake. Therefore, one of the most commonmethods for weight control to combat obesity is the use of relativelylow-fat diets, that is, diets containing less fat than a “normal diet”or that amount usually consumed by the patient.

[0003] The presence of fats in a great many food sources greatly limitsthe food sources which can be used in a low fat diet. Additionally, fatscontribute to the flavor, appearance and physical characteristics ofmany foodstuffs. As such, the acceptability of low-fat diets and themaintenance of such diets are difficult.

[0004] Various chemical approaches have been proposed for controllingobesity. Anorectic agents such as dextroamphetamine, the combination ofthe non-amphetamine drugs phentermine and fenfluramine (Phen-Fen), anddexfenfluramine (Redux) alone, are associated with serious side effects.Indigestible materials such as olestra (OLEAN®), mineral oil orneopentyl esters (see U.S. Pat. No. 2,962,419) have been proposed assubstitutes for dietary fat. Garcinia acid and derivatives thereof havebeen described as treating obesity by interfering with fatty acidsynthesis. Swellable crosslinked vinyl pyridine resins have beendescribed as appetite suppressants via the mechanism of providingnon-nutritive bulk, as in U.S. Pat. No. 2,923,662. Surgical techniquessuch as temporary ileal bypass surgery, are employed in extreme cases.

[0005] However, methods for treating obesity, such as those describedabove have serious shortcomings with controlled diet remaining the mostprevalent technique for controlling obesity. As such, new methods fortreating obesity are needed.

SUMMARY OF THE INVENTION

[0006] The invention features a method for treating obesity in a patientby administering to the patient a polymer that has been substituted withor comprises one or more groups which can inhibit a lipase. Lipases arekey enzymes in the digestive system which break down tri- anddiglycerides, which are too large to be absorbed by the small intestineinto fatty acids which can be absorbed. Therefore, inhibition of lipasesresults in a reduction in the absorption of fat. In one embodiment, thelipase inhibiting group can be a “suicide substrate” which inhibits theactivity of the lipase by forming a covalent bond with the enzyme eitherat the active site or elsewhere. In another embodiment, the lipaseinhibiting group is an isosteric inhibitor of the enzyme. The inventionfurther relates to the polymers employed in the methods described hereinas well as novel intermediates and methods for preparing the polymers.

DETAILED DESCRIPTION OF THE INVENTION

[0007] The invention features a method for treating obesity in a patientby administering to the patient a polymer comprising one or more groupswhich can inhibit a lipase. Since lipases are responsible for thehydrolysis of fat, a consequence of their inhibition is a reduction infat hydrolysis and absorption. The invention further relates to thepolymers employed in the methods described herein as well as novelintermediates and methods for preparing the polymers.

[0008] In one aspect of the invention, the lipase inhibiting groupinactivates a lipase such as gastric, pancreatic and lingual lipases.Inactivation can result by forming a covalent bond such that the enzymeis inactive. The covalent bond can be formed with an amino acid residueat or near the active site of the enzyme, or at a residue which isdistant from the active site provided that the formation of the covalentbond results in inhibition of the enzyme activity. Lipases contain acatalytic triad which is responsible for the hydrolysis of lipids intofatty acids. The catalytic triad consists of a serine, aspartate andhistidine amino acid residues. This triad is also responsible for thehydrolysis of amide bonds in serine proteases, and it is expected thatcompounds that are serine protease inhibitors will also inhibit lipases.Therefore, serine protease inhibitors that can be covalently linked to apolymer are preferred lipase inhibiting groups. For example, a covalentbond can be formed between the lipase inhibiting group and a hydroxyl ator the catalytic site of the enzyme. For instance, a covalent bond canbe formed with serine. Inactivation can also result from a lipaseinhibiting group forming a covalent bond with an amino acid, for examplecysteine, which is at some distance from the active site. In addition,non-covalent interaction between the lipase inhibiting group and theenzyme can also result in inactivation of the enzyme. For example, thelipase inhibiting group can be an isostere of a fatty acid, which caninteract non-covalently with the catalytic site of the lipase. Inaddition, the lipase inhibiting group can compete for lipase hydrolysiswith natural triglycerides.

[0009] In one aspect of the invention, a lipase inhibiting group can berepresented by formula I:

[0010] wherein,

[0011] R is a hydrogen, hydrophobic moiety, —NR²R³, —CO₂H, —OCOR²,—NHCOR², a substituted or unsubstituted aliphatic group or a substitutedor unsubstituted aromatic group;

[0012] R¹ is an activating group;

[0013] Y is oxygen, sulfur, —NR²— or is absent;

[0014] Z and Z¹ are, independently, an oxygen, alkylene, sulfur, —SO₃—,—CO₂—, —NR²—, —CONR²—, —PO₄H— or a spacer group;

[0015] R² and R³ are, independently, a hydrogen, a substituted orunsubstituted aliphatic group, or a substituted or unsubstitutedaromatic group;

[0016] m is 0 or 1; and

[0017] n is 0 or 1.

[0018] In one embodiment, the lipase inhibiting group of formula I canbe represented by the following structures:

[0019] wherein R, R¹ and Y are defined as above.

[0020] In another embodiment, the lipase inhibiting group of structuralformula I can be represented by the following structures:

[0021] wherein R, R¹, R², R³ and Y are defined as above, and p is aninteger (e.g. an integer between zero and about 30, preferably betweenabout 2 and about 10).

[0022] In another embodiment, the lipase inhibitor of formula I is amixed anhydride. Mixed anhydrides include, but are not limited to,phosphoric-carboxylic, phosphoric-sulfonic and pyrophosphate mixedanhydride lipase inhibiting groups which can be represented by thefollowing structures, respectively:

[0023] wherein R, R¹, Y and Z¹ are defined as above.

[0024] In another aspect, a lipase inhibiting group of the invention canbe an anhydride. In one embodiment, the anhydride is a cyclic anhydriderepresented by formula II:

[0025] wherein R, Z and p are defined as above, X is —PO₂—, —SO₂— or—CO—, and k is an integer from 1 to about 10, preferably from 1-4.

[0026] In another embodiment, the anhydride lipase inhibiting groups canbe a cyclic anhydride which is part of a fused ring system. Anhydridesof this type can be represented by formula III:

[0027] wherein X and Z are defined as above, and ring A is an optionallysubstituted cyclic aliphatic group or aromatic group, or combinationsthereof, which can include one or more heteroatoms in the ring. In aparticular embodiment, the cyclic anhydride is a benzenesulfonicanhydride represented by the following structure:

[0028] wherein Z is defined as above and the benzene ring can be furthersubstituted.

[0029] In another aspect, the lipase inhibiting group is anα-halogenated carbonyl which can be represented by formula IV:

[0030] wherein R and Y are defined as above, and W¹ and W² are eachindependently hydrogen or halogen, for example, —F, —Cl, —Br, and —I,wherein at least one of W¹ and W² is a halogen.

[0031] In yet another aspect, a cyclic compound having an endocyclicgroup that is susceptible to nucleophilic attack can be a lipaseinhibiting group. Lactones and epoxides are examples of this type oflipase inhibiting group and can be represented by formulas V and VI,respectively:

[0032] wherein R, Z, m and p are defined as above.

[0033] In a further aspect, the lipase inhibiting group can be asulfonate or disulfide group represented by formulas VII and VIII,respectively:

[0034] wherein R, Z and p are defined as above, and R⁵ is absent or ahydrophobic moiety, a substituted or unsubstituted aliphatic group or asubstituted or unsubstituted aromatic group.

[0035] In a particular embodiment, the disulfide lipase inhibiting groupcan be represent by the following formula:

[0036] wherein R, Z and p are defined as above.

[0037] In a further aspect of the invention, a lipase inhibiting groupcan be a boronic acid which can be linked to a polymer by a hydrophobicgroup or to the polymer directly when the polymer is hydrophobic.Boronic acid lipase inhibiting groups can be represented by thefollowing structure:

[0038] wherein R⁵, Z, n and m are defined as above.

[0039] In an additional aspect, an isosteric lipase inhibiting group canbe a phenolic acid linked to the polymer. Phenolic acid lipaseinhibiting groups can be represented by the following structure:

[0040] wherein Z, R⁵, n and m are defined as above and —CO₂H and —OH areortho or para with respect to each other.

[0041] A variety of polymers can be employed in the invention describedherein. The polymers can be aliphatic, alicyclic or aromatic orsynthetic or naturally occurring. However, aliphatic and alicyclicsynthetic polymers are preferred. Furthermore, the polymer can behydrophobic, hydrophilic or copolymers of hydrophobic and/or hydrophilicmonomers. The polymer can be non-ionic (e.g., neutral), anionic orcationic, in whole or in part. Furthermore, the polymers can bemanufactured from olefinic or ethylenic monomers (such as vinylalcohol)or condensation polymers.

[0042] For example, the polymers can be a polyvinylalcohol,polyvinylamine, poly-N-alkylvinylamine, polyallylamine,poly-N-alkylallylamine, polyalkylenimine, polyethylene, polypropylene,polyether, polyethylene oxide, polyamide, polyacrylic acid,polyalkylacrylate, polyacrylamide, polymethacrylic acid,polyalkylmethacrylate, polymethacrylamide, poly-N-alkylacrylamide,poly-N-alkylmethacrylamide, polystyrene, vinylnaphthalene,ethylvinylbenzene, aminostyrene, vinylbiphenyl, vinylanisole,vinylimidazolyl, vinylpyridinyl, dimethylaminomethylstyrene,trimethylammoniumethylmethacrylate, trimethylammoniumethylacrylate,carbohydrate, protein and substituted derivatives of the above (e.g.,fluorinated monomers thereof and copolymers thereof.

[0043] Preferred polymers include polyethers, such as polyalkyleneglycols. Polyethers can be represented by the formula IX:

[0044] wherein R is defined as above and q is an integer.

[0045] For example, the polymer can be polypropylene glycol orpolyethylene glycol or copolymers thereof. The polymers can be random orblock copolymers. Also, the polymers can be hydrophobic, hydrophilic, ora combination thereof (as in random or block polymers).

[0046] A particularly preferred polymer is a block copolymercharacterized by hydrophobic and hydrophilic polymeric regions. In suchan embodiment, the “core polymer can be hydrophobic with one or bothends capped with a hydrophilic polymer or vice versa. An example of sucha polymer is a polyethyleneglycol-polypropyleneglycol-polethyleneglycolcopolymer, as is sold under the tradename PLURONIC® (BASF WyandotteCorp.). BRIJ® and IGEPAL® (Aldrich, Milwaukee, Wis.) are examples ofpolymers having a polyethylene glycol core capped withe a hydrophobicend group. BRIJ® polymers are polyethylene glycols having one end cappedwith alkoxy group, while the hydroxy group at the other end of thepolymer chain is free. IGEPAL® polymers are polyethylene glycols havingone end capped with 4-nonylphenoxy group, while the hydroxy group at theother end of the polymer chain is free.

[0047] Another class of polymers includes aliphatic polymers such as,polyvinylalcohol, polyallylamine, polyvinylamine and polyethylenimine.These polymers can be further characterized by one or more substituents,such as substituted or unsubstituted, saturated or unsaturated alkyl andsubstituted or unsubstituted aryl. Suitable substituents includeanionic, cationic or neutral groups, such as alkoxy, aryl, aryloxy,aralkyl, halogen, amine, and ammonium groups, for example. The polymercan desirably possess one or more reactive functional groups which can,directly or indirectly, react with an intermediate possessing the lipaseinhibiting groups.

[0048] In one embodiment, the polymers have the following repeat unit:

[0049] wherein,

[0050] q is an integer; and

[0051] R⁴ is —OH, —NH₂, —CH₂NH₂, —SH, or a group represented by thefollowing formula:

[0052] wherein R, R¹, Y, Z, Z¹, m and n are defined as above.

[0053] Additionally, the polymer can be a carbohydrate, such aschitosan, cellulose, hemicellulose or starch or derivatives thereof.

[0054] The polymer can be linear or crosslinked. Crosslinking can beperformed by reacting the copolymer with one or more crosslinking agentshaving two or more functional groups, such as electrophilic groups,which react with an alcohol of the polymer to form a covalent bond.Crosslinking in this case can occur, for example, via nucleophilicattack of the polymer hydroxy groups on the electrophilic groups. Thisresults in the formation of a bridging unit which links two or morealcoholic oxygens from different polymer strands. Suitable crosslinkingagents of this type include compounds having two or more groups selectedfrom among acyl chloride, epoxide, and alkyl-X, wherein X is a suitableleaving group, such as a halo, tosyl or mesyl group. Examples of suchcompounds include, but are not limited to, epichlorohydrin, succinyldichloride, acryloyl chloride, butanedioldiglycidyl ether,ethanedioldiglycidyl ether, pyromellitic dianhydride, and dihaloalkanes.

[0055] The polymer composition can also be crosslinked by including amultifunctional co-monomer as the crosslinking agent in the reactionmixture. A multifunctional co-monomer can be incorporated into two ormore growing polymer chains, thereby crosslinking the chains. Suitablemultifunctional co-monomers include, but are not limited to,diacrylates, triacrylates, and tetraacrylates, dimethacrylates,diacrylamides, diallylacrylamides, and dimethacrylamides. Specificexamples include ethylene glycol diacrylate, propylene glycoldiacrylate, butylene glycol diacrylate, ethylene glycol dimethacrylate,butylene glycol dimethacrylate, methylene bis(methacrylamide), ethylenebis(acrylamide), ethylene bis(methacrylamide), ethylidenebis(acrylamide), ethylidene bis(methacrylamide), pentaerythritoltetraacrylate, trimethylolpropane triacrylate, bisphenol Adimethacrylate, and bisphenol A diacrylate. Other suitablemultifunctional monomers include polyvinylarenes, such asdivinylbenzene.

[0056] The molecular weight of the polymer is not critical. It isdesirable that the polymer be large enough to be substantially orcompletely non-absorbed in the GI tract. For example, the molecularweight can be more than 900 Daltons.

[0057] The digestion and absorption of lipids is a complex process inwhich water insoluble lipids are emulsified to form an oil in wateremulsion with an oil droplet diameter of approximately 0.5 mm. Thisemulsified oil phase has a net negative charge due to the presence offatty acids and bile salts, which are the major emulsifying agents.Lipases that are present in the aqueous phase hydrolyze the emulsifiedlipids at the emulsion surface. Most lipases contain an active site thatis buried by a surface loop of amino acids that sit directly on top ofthe active site when the lipase is in an aqueous solution. However, whenthe lipase comes in contact with bile salts at the lipid/water interfaceof a lipid emulsion, the lipase undergoes a conformational change thatshifts the surface loop to one side and exposes the active site. Thisconformational change allows the lipase to catalyze hydrolysis of lipidsat the lipid/water interface of the emulsion. Polymers that disrupt thesurface of the emulsion or alter its chemical nature are expected toinhibit lipase activity. Therefore, it may increase the effectiveness ofpolymers that have lipase inhibiting groups to administer them with oneor more polymers that alter the emulsion surface. Alternatively, lipaseinhibiting groups can be attached directly to such a polymer.

[0058] Several types of fat-binding polymers have been effective indisrupting the surface of the lipid emulsion or altering its chemicalnature. For example, polymers that have positively charged emulsifiersare able to form stable polycation lipid emulsions. The lipids in suchan emulsion are not substrates for gastrointestinal lipases because thesurface of the emulsion has a net positive charge instead of the usualnet negative charge. Another type of fat-binding polymer destabilizesthe emulsion causing the oil droplets of the emulsion to coalesce. Thisdecreases the emulsion surface area where lipases are active, andtherefore, reduces lipid hydrolysis. Fat-binding polymer are furtherdefined in copending application Ser. No. 09/004,963, filed on Jan. 9,1998, and application Ser. No. 09/166,453, filed on Oct. 5, 1998, thecontents of which are incorporated herein by reference.

[0059] The substituted polymers described herein can be manufacturedaccording to methods generally known in the art. For example, a lipaseinhibiting intermediate characterized by a reactive moiety can becontacted with a polymer characterized by a functional group whichreacts with said reactive moiety. See March, J., Advanced OrganicChemistry, 3^(rd) edition, John Wiley and Sons, Inc.; New York, (1985).

[0060] A “hydrophobic moiety,” as the term is used herein, is a moietywhich, as a separate entity, is more soluble in octanol than water. Forexample, the octyl group (C₈H₁₇) is hydrophobic because its “parent”alkane, octane, has greater solubility in octanol than in water. Thehydrophobic moieties can be a saturated or unsaturated, substituted orunsubstituted hydrocarbon group. Such groups include substituted andunsubstituted, normal, branched or cyclic aliphatic groups having atleast four carbon atoms, substituted or unsubstituted arylalkyl orheteroarylalkyl groups and substituted or unsubstituted aryl orheteroaryl groups. Preferably, the hydrophobic moiety includes analiphatic group of between about six and thirty carbons. Specificexamples of suitable hydrophobic moieties include the following alkylgroups: butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl,tetradecyl, hexadecyl, octadecyl, docosanyl, cholesteryl, farnesyl,aralkyl, phenyl, and naphthyl, and combinations thereof. Other examplesof suitable hydrophobic moieties include haloalkyl groups of at leastfourcarbons (e.g., 10-halodecyl), halodecyl), hydroxyalkyl groups of atleast six carbons (e.g., 11-hydroxyundecyl), and aralkyl groups (e.g.,benzyl). As used herein aliphatic groups include straight, chained,branched or cyclic C₄-C₃₀ hydrocarbons which are completely saturated orcontain one or more units of unsaturation.

[0061] Aromatic groups suitable for use in the invention include, butare not limited to, aromatic rings, for example, phenyl and substitutedphenyl, heteroaromatic rings, for example, pyridinyl, furanyl andthiophenyl, and fused polycyclic aromatic ring systems in which acarbocyclic aromatic ring or heteroaryl ring is fused to one or moreother carbocyclic or heteroaryl rings. Examples of fused polycyclicaromatic ring systems include substituted or unsubstituted phenanthryl,anthracyl, naphthyl, 2-benzothienyl, 3-benzothienyl, 2-benzofuranyl,3-benzofuranyl, 2-indolyl, 3-indolyl, 2-quinolinyl, 3-quinolinyl,2-benzothiazole, 2-benzooxazole, 2-benzimidazole, 2-quinolinyl,3-quinolinyl, 1-isoquinolinyl, 3-quinolinyl, 1-isoindolyl, 3-isoindolyl,and acridintyl.

[0062] A “substituted aliphatic or aromatic group” can have one or moresubstituents, e.g., an aryl group (including a carbocyclic aryl group ora heteroaryl group), a substituted aryl group, —O-(aliphatic group oraryl group), —O-(substituted aliphatic group or substituted aryl group),acyl, —CHO, —CO-(aliphatic or substituted aliphatic group), —CO-(aryl orsubstituted aryl), —COO-(aliphatic or substituted aliphatic group),—COO-(aryl or substituted aryl group), —NH-(acyl), —O-(acyl), benzyl,substituted benzyl, halogenated lower alkyl (e.g. trifluoromethyl andtrichloromethyl), fluoro, chloro, bromo, iodo, cyano, nitro, —SH,—S-(aliphatic or substituted aliphatic group), —S-(aryl or substitutedaryl), —S-(acyl) and the like.

[0063] An “activating group” is a group that renders a functional groupor moiety reactive. Generally, electron withdrawing groups are“activating groups.” R¹ or Y—R¹, of the above formulae, is preferably agood leaving group or an electron withdrawing group. Examples of goodleaving groups are phosphate, p-nitrophenol, o,p-dinitrophenol,N-hydroxysuccinimide, imidazole, ascorbic acid, pyridoxine,trimethylacetate, adamantanecarbonylate, p-chlorophenol,o,p-dichlorophenol, methanesulfonylate, mesitylsulfonylate andtriisopropylbenzenesulfonylate. A preferred leaving group isN-hydroxysuccinimide.

[0064] A spacer group can be a group that has one to about thirty atomsand is covalently bonded to the lipase inhibitor, to the polymer, or tothe hydrophobic moiety. Generally, the spacer group can be covalentlybonded to the lipase inhibitor, polymer or hydrophobic moiety through afunctional group. Examples of functional groups are oxygen, alkylene,sulfur, —SO₂—, —CO₂—, —NR²—, or —CONR²—. A spacer group can behydrophilic or hydrophobic. Examples of spacer groups include aminoacids, polypeptides, carbohydrates, and optionally substituted alkyleneor aromatic groups. Spacer groups can be manufactured from, for example,epichlorohydrin, dihaloalkane, haloalkyl esters, polyethylene glycol,polypropylene glycol and other cross-linking or difunctional compounds.Bromoalkylacetate is a preferred spacer group.

[0065] The amount of a polymer administered to a subject will depend onthe type and severity of the disease and on the characteristics of thesubject, such as general health, age, body weight and tolerance todrugs. It will also depend on the degree of obesity and obesity relatedcomplications. The skilled artisan will be able to determine appropriatedosages depending on these and other factors. Typically, in humansubjects, an effective amount of the polymer can range from about 10 mgper day to about 50 mg per day for an adult. Preferably, the dosageranges from about 10 mg per day to about 20 mg per day.

[0066] The polymer can be administered by any suitable route, including,for example, orally in capsules, suspensions or tablets. Oraladministration by mixing with food is a preferred mode ofadministration.

[0067] The polymer can be administered to the individual in conjunctionwith an acceptable pharmaceutical carrier as part of a pharmaceuticalcomposition. Formulation of a polymer to be administered will varyaccording to the route of administration selected (e.g., solution,emulsion, capsule). Suitable pharmaceutical carriers may contain inertingredients which do not interact with the lipase inhibiting groups ofthe polymer. Standard pharmaceutical formulation techniques can beemployed, such as those described in Remington's PharmaceuticalSciences, Mack Publishing Company, Easton, Pa. Methods for encapsulatingcompositions (such as in a coating of hard gelatin or cyclodextran) areknown in the art (Baker, et al., “Controlled Release of BiologicalActive Agents”, John Wiley and Sons, 1986).

[0068] Experimental

[0069] Synthesis of Polymers

EXAMPLE 1

[0070] Preparation of polyethylene glycol having an n-pentyl hydroprobicmoiety and p-nitrophenyl phosphate lipase inhibiting groups:

[0071] A mixture of n-pentanol (19.5 mmol. 1.72 g) and N-methylimidazole (19.5 mmol, 1.6 g) in anhydrous methylene chloride (40 mL) wasadded slowly over 20 minutes under anhydrous conditions to a solution ofp-nitrophenyl phosphorodichloridate (5.0 g, 19.5 mmol) in anhydrousmethylene chloride (100 mL). The reaction flask was cooled in a waterbath during the addition. After the completion of the addition, thewater bath was removed, and the reaction mixture was stirred for 2 hoursat room temperature. A mixture of polyethyleneglycol (MW=8,000; 10 mmol,80 g), and N-methyl imidazole (19.5 mmol, 1.6 g) in anhydrous methylenechloride (150 mL) was added to the reaction flask under anhydrousconditions. The mixture was stirred for 25 hours at room temperature.The solvent was removed under vacuum, the residue was purified accordingto method A, and the polymer was obtained as white powder (70 g).

[0072] Purification Procedures

[0073] Method A:

[0074] The residue was dissolved in de-ionized water (100 mL). Thesolution was dialyzed for 24 hours using Spectra/Por Membrane MWCO:3,500. The dialyzed solution was lyophilized, and the polymer wasobtained as white powder.

[0075] Method B:

[0076] The residue was poured into 0.5 L of diethyl ether and stirred atroom temperature for 1 hour. The solvent was decanted and replaced withfresh diethyl ether (0.25 L). The mixture was stirred for 1 hour. Thesolvent was removed, and the polymer was dried at room temperature undervacuum.

[0077] Method C:

[0078] The reaction mixture was washed with 10% aqueous sodium sulfatesolution (3×100 mL). The organic phase was dried over magnesium sulfate.The solvent was removed, and the polymer was dried at room temperature.

[0079] Using the above procedures, the following compounds weresynthesized and are tabulated in the following table. TABLE 1Polyethylene glycols (PEG) having p-nitrophenyl phosphate lipaseinhibiting groups with a variety of hydroprobic moieties. HYDROPHOBICPEG MOIETY METHOD OF PHYSICAL EXAMPLE MW (R) PURIFICATION STATE 1 8,400n-pentyl Method A powder 2 3,400 n-decyl Method B powder 3 3,400n-dodecyl Method B powder 4 3,400 n-octadecyl Method B powder 5 1,000n-decyl Method B semi solid 6 1,000 n-dodecyl Method B semi solid 71,000 n-tetradecyl Method B semi solid 8 1,000 n-hexadecyl Method B semisolid 9 1,000 n-octadecyl Method B semi solid 10 1,000 n-pentyl Method Csemi solid 11 1,000 n-hexyl Method C semi solid 12 1,000 n-octyl MethodC semi solid 13 1,000 n-docosyl Method C powder 14 1,000 cholesterylMethod C powder 15 3,400 n-pentyl Method B solid 16 1,500 n-pentylMethod B solid 17 1,500 n-decyl Method B solid 18 1,500 n-dodecyl MethodB solid 19 1,500 n-hexadecyl Method C solid 20 1,500 n-octadecyl MethodC solid 21 1,500 n-docosyl Method B solid 22 1,500 rac-farnesyl Method Bbrown, solid 23 1,500 n-cholesteryl Method C solid 24 1,5005-phenyl-1-pentyl Method C solid 25 1,500 n-octyl Method C solid 261,500 n-hexyl Method C solid 27 3,400 n-octyl Method C solid 28 8,400n-octyl Method C solid

EXAMPLE 29

[0080] Preparation of a PLURONIC® polymer having a n-tetradecylhydrophobic moiety and p-nitrophenyl phosphate lipase inhibiting groups:

[0081] A mixture of n-tetradecanol (15 g, 70 mmol) and N-methylimidazole (5.6 mL, 70 mmol) in anhydrous methylene chloride (75 mL) wasadded slowly over 20 minutes under anhydrous condition to a solution ofp-nitrophenyl phosphorodichloridate (17.92 g, 70 mmol) in anhydrousmethylene chloride (50 mL). The reaction flask was cooled in a waterbath during the addition. After the completion of the addition, thewater bath was removed, and the reaction mixture was stirred for 2 hoursat room temperature. A mixture of PLURONIC® (MW=1,100; 39 g, 35 mmol)and N-methyl imidazole (5.6 mL. 70 mmol) in anhydrous methylene chloride(150 mL) was added to the reaction flask under anhydrous conditions. Themixture was stirred for 24 hours at room temperature. The reactionmixture was extracted with cold saturated NaCl solution (3×150 mL), theorganic layer was dried over anhydrous sodium sulfate. The sodiumsulfate was removed by filtration, and the filtrate was collected. Thesolvent was removed from the filtrate under reduced pressure to give 65g of pale yellow colored viscous liquid. The material was dried undervacuum for one week at room temperature. This was used directly for thein vitro and in vivo assay.

[0082] The following Examples were prepared using the above procedure.TABLE 2 PLURONIC ® Polymers (PLU) having p-nitrophenyl phosphate lipaseinhibiting groups with a variety of hydrophobic moieties. WT. %HYDROPHOBIC PLU ETHYLENE MOIETY PHYSICAL EXAMPLE MW GLYCOL (R) STATE 291,100 10 wt % n-tetradecyl liquid 30 1,100 10 wt % n-dodecyl liquid 311,100 10 wt % n-decyl liquid 32 1,100 10 wt % n-oetyl liquid 33 1,900 50wt % n-hexyl liquid 34 1,900 50 wt % n-octyl liquid 35 1,900 50 wt %n-decyl liquid 36 1,900 50 wt % n-dodecyl liquid 37 1,900 50 wt %n-tetradecyl semi solid 38 1,900 50 wt % n-hexadecyl semi solid 39 8,40080 wt % n-pentyl powder 40 8,400 80 wt % n-hexyl powder 41 2,900 40 wt %n-octadecyl semi solid 42 2,900 40 wt % n-hexadecyl semi solid 43 2,90040 wt % n-tetradecyl liquid 44 2,900 40 wt % n-dodecyl liquid 45 4,40040 wt % n-octadecyl semi solid 46 4,400 40 wt % n-hexadecyl semi solid47 4,400 40 wt % n-tetradecyl liquid 48 4,400 40 wt % n-dodecyl liquid

EXAMPLE 51

[0083] Preparation of a polypropylene glycol having a n-hexadecylhydrophobic moiety and p-nitrophenyl phosphate lipase inhibiting group:

[0084] A mixture of n-hexadecanol (28.41 g, 117 mmol) and N-methylimidazole (9.34 mL, 117 mmol) in anhydrous methylene chloride (75 mL)was added slowly over 20 minutes under anhydrous condition to a solutionof p-nitrophenyl phosphorodichloridate (30 g, 117 mmol) in anhydrousmethylene chloride (60 mL). The reaction flask was cooled in a waterbath during the addition. After the completion of the addition, thewater bath was removed and the reaction mixture was stirred for 2 hoursat room temperature. A mixture of polypropylene glycol (MW=1000; 58.5 g,58.5 mmol) and N-methyl imidazole (9.3 mL, 117 mmol) in anhydrousmethylene chloride (150 mL) was added to the reaction flask underanhydrous conditions. The mixture was stirred for 24 hours at roomtemperature. The reaction mixture was extracted with cold saturatedsolution of Na₂SO₄ (3×150 mL). The organic layer was dried overanhydrous magnesium sulfate. The magnesium sulfate was removed byfiltration, and the filtrate was collected. The solvent was removed fromthe filtrate under reduced pressure to give a product of 77 g. Thematerial was dried under vacuum at room temperature for 4 days.

[0085] The following polypropylene glycol p-nitrophenyl phosphates wereprepared using the above procedure. TABLE 3 Polypropylene glycol (PPG)having p-nitrophenyl phosphate lipase inhibiting groups with a varietyof hydrophobic moieties. HYDROPHOBIC PPG MOIETY PHYSICAL EXAMPLE MW (R)STATE 49 1,000 n-pentyl semi solid 50 1,000 n-octyl semi solid 51 1,000n-hexadecyl semi solid 52 1,000 n-octadecyl semi solid 53 2,000 n-pentylsemi solid 54 2,000 n-octyl semi solid 55 2,000 n-hexadecyl semi solid56 2,000 n-octadecyl semi solid

EXAMPLE 57

[0086] Preparation of a polyethylene glycol polymer having ap-nitrophenyl phosphonate lipase inhibiting group and having a pentylhydrophobic moieties:

[0087] A. Preparation of O,O-dimethyl n-pentylphosphonate:

[0088] O,O-dimethyl phosphonate (220 g, 2 mol) was added dropwise to asuspension of NaH (48 g, 2 mol) in anhydrous THF (600 mL) undernitrogen. After 1 hour, 1-bromopentane (248 mL, 2 mol) in THF (400 mL)was added slowly, and the reaction mixture was refluxed for 12 hours.The solvent was removed under vacuum, diethyl ether (1 L) was added, andthe salts were removed by filtration. The ether solution was washed withwater (3×100 mL), the organic layer was dried over anhydrous sodiumsulfate. The ether was removed under reduced pressure, and the crudeproduct was purified by distillation under vacuum to give 171 g ofO,O-dimethyl n-pentyl phosphonate.

[0089] B. Preparation of n-pentylphosphonic dichloride:

[0090] O,O-dimethyl n-pentyl phosphonate (158 g, 0.88 mol) andN,N-dimethyl formamide (700 mg) were dissolved in thionyl chloride (200mL), and the resulted mixture was refluxed for 48 hours. The volatileswere removed under vacuum at room temperature, and the crude product waspurified by distillation to give a colorless liquid (135 g).

[0091] C. Preparation of polyethylene glycol Having a p-nitrophenyln-pentyl phosphonate lipase Inhibiting Groups:

[0092] To a solution of n-pentylphosphonic dichloride (2.65 g, 14 mmol)in 40 mL of anhydrous dichloromethane, was added bright orange coloredsodium salt of p-nitrophenol (2.3 g, 14 mmol) under anhydrous condition.The bright orange color disappeared within 5-10 minutes. After 45minutes, a mixture of polyethylene glycol (MW=8,400; 56 g, 7 mmol) andN-methylimidazole (1.5 mL, 20 mmol) was added at room temperature andstirred for 24 hours. The reaction mixture was washed with 2% K₂CO₃solution (6×100 mL) followed by saturated NaCl solution (6×100 mL). Theorganic layer was dried over Na₂SO₄, the solvent was removed undervacuum to give a viscous liquid. The product was poured into 200 mL ofdiethyl ether and stirred for 10 minutes. The ether portion wasdecanted, and the procedure was repeated three more times. The productwas obtained as a white powder which was dried under vacuum at roomtemperature for a week.

[0093] The following polyethylene glycol polymers having p-nitrophenylphosphonate lipase inhibiting groups were prepared by this procedure.TABLE 4 Polyethylene glycols having p-nitrophenyl phosphonate lipaseinhibiting groups with a pentyl hydrophobic moiety. HYDROPHOBIC MOIETYPHYSICAL EXAMPLE PEG (R) STATE 57 8,400 n-pentyl powder 58 3,400n-pentyl powder 59 1,500 n-pentyl semi solid 60 1,000 n-pentyl semisolid

EXAMPLE 61

[0094] Preparation of a PLURONIC® polymer having p-nitrophenylphosphates lipase inhibiting group tethered by a n-pentyl-1,5-dioxylinker and having a n-hexadecyl hydrophobic moiety:

[0095] A 1 L, round-bottomed flask was charged with sodium hydride (4.0g as a 60% dispersion of NaH in mineral oil, 0.1 mol) then washed withanhydrous heptane (3×25 mL). Anhydrous tetrahydrofuran (THF) (150 mL)was added, and the suspension was stirred at room temperature undernitrogen. A solution of PLURONIC® (MW=1900, 50 wt % polyethylene glycol,50 wt % polypropylene glycol; 95 g, 0.05 mole) in anhydrous THF (200 mL)was added at room temperature. A solution of bromopentyl acetate (20.9g, 0.1 mole) in anhydrous THF (50 mL) was added to the reaction mixtureunder anhydrous conditions. The reaction mixture was refluxed at 60° C.for 16 hours. The solvent was removed under vacuum, and the resultingslurry was suspended in dichloromethane (300 mL). The solids wereremoved by filtration, and the filtrate was washed with water (3×100mL). The organic layer was dried over anhydrous sodium sulfate, and thesolvent was removed to give a pale brown viscous liquid (110 g). Thismaterial was dissolved in methanol (500 mL) and treated with aqueous 4NNaOH (40 mL). After 4 hours, the reaction mixture was acidified withconcentrated HCl, and the solvent was removed under vacuum. The viscousoil was dissolved in dichloromethane, which was washed with water (4×100mL). The organic layer was dried over sodium sulfate, and the solventwas removed to give bis-5-hydroxypentoxy PLURONIC® as a pale brownviscous liquid (98 g).

[0096] In a separate flask, a mixture of n-hexadecanol (7.02 g, 29.0mmol) and N-methyl imidazole (2.3 mL, 290 mmol) in anhydrous methylenechloride (40 mL) was added slowly over 20 minutes under anhydrousconditions to a solution of p-nitrophenyl phosphorodichloridate (7.41 g,29.0 mmol) in anhydrous methylene chloride (100 mL). The reaction flaskwas cooled in a water bath during the addition. After the completion ofthe addition, the water bath was removed, and the reaction mixture wasstirred for 2 hours at room temperature. A mixture ofbis-5-hydroxypentoxy PLURONIC® (30 g, 14.48 mmol), and N-methylimidazole (2.3 mL) in anhydrous methylene chloride (150 mL) was added tothe reaction flask under anhydrous conditions. The mixture was stirredfor 24 hours at room temperature, then washed with saturated NaClsolution (3×100 mL). The organic layer was collected and dried oversodium sulfate. The solvent was removed to give a viscous liquid. Thiswas washed with boiling hexane (6×50 mL), and the product was driedunder vacuum at room temperature overnight to yield a pale yellowviscous liquid (39 g).

[0097] The following Examples were prepared using the above procedure.TABLE 5 PLURONIC ® polymers having p-nitrophenyl phosphate lipaseinhibiting groups tethered by a variety of dialkoxys and having avariety of hydrophobic moieties. HYDROPHOBIC PLU MOIETY DIALKOXY EXAMPLEMW (R) (Z¹) 61 1900 n-pentyl n-pent-1,5-dioxy 62 1900 n-decyln-pent-1,5-dioxy 63 1900 n-hexadecyl n-pent-1,5-dioxy 64 1900 n-pentyln-undecyl-1,10-dioxy 65 1900 n-decyl n-undecyl-1,10-dioxy 66 1900n-hexadecyl n-undecyl-1,10-dioxy

EXAMPLE 67

[0098] Preparation of a polyethylene glycol polymer having ap-nitrophenyl phosphates lipase inhibiting group tethered by an-pentyl-1,5-dioxy linker and having a n-hexadecyl hydrophobic moiety:

[0099] A 1 L, round-bottomed flask was charged with sodium hydride (7.67g as a 60% dispersion of NaH in mineral oil, 0.19 mol) and was washedwith anhydrous heptane (3×25 mL). Anhydrous THF (200 mL) was added, andthe suspension was stirred at room temperature under nitrogen. Asolution of polyethylene glycol (MW=1,500; 150 g, 0.1 mol) in anhydrousTHF (200 mL) was added at room temperature under anhydrous conditions.The mixture was stirred for 1 hour at room temperature, then a solutionof bromopentyl acetate (41.82 g, 0.2 mol) in anhydrous THF (100 mL) wasadded to the reaction mixture. The reaction mixture was refluxed at 60°C. for 16 hours. The solvent was removed under vacuum, and the resultingslurry was suspended in dichloromethane (300 mL). The solids wereremoved by filtration, and the filtrate was washed with water (3×100mL). The organic layer was dried over anhydrous sodium sulfate, and thesolvent was removed to give a pale brown viscous liquid (110 g). Thismaterial was dissolved in methanol (500 mL) and treated with aqueous 4NNaOH (80 mL). After 4 hours, the reaction mixture was acidified withconcentrated HCl, and the solvent was removed under vacuum. The viscousoil was dissolved in dichloromethane and was washed with water (4×100mL). The organic layer was dried over sodium sulfate, and the solventwas removed to give a bis-5-hydroxypentoxy polyethylene glycol as a palebrown viscous liquid (98 g). The p-nitrophenyl phosphate group was addedin a manner analogous to the procedure in Example 61.

[0100] The following Examples were prepared using the above procedure.TABLE 6 Polyethylene glycols having a p-nitrophenyl phosphate lipaseinhibiting group tethered by a dialkoxy linker and having a variety ofhydrophobic moieties. HYDROPHOBIC PEG MOIETIES DIALKOXY EXAMPLE MW (R)(Z¹) 67 1500 n-hexyl n-pent-1,5-dioxy 68 1500 n-dodecyl n-pent-1,5-dioxy69 1500 n-hexadecyl n-pent-1,5-dioxy

EXAMPLE 75

[0101] Preparation of a BRIJ® polymer having a p-nitrophenyl phosphatelipase inhibiting group and a hexadecyl hydrophobic moiety:

[0102] p-Nitrophenyl phosphorodichloridate (75 g, 0.29 mol) in anhydrousdichloromethane (300 mL) was added to a 1 L, three necked,round-bottomed flask with stir bar that had been purged with N₂. Asolution of hexadecanol (71.03 g, 0.29 mol) and N-methylimidazole (23.35mL, 0.29 mol) in anhydrous dichloromethane (250 mL) was added dropwiseover a period of 2 hours. The reaction mixture was stirred for anadditional 1 hour before pouring into a 1 L separatory funnel.N-methylimidazole hydrochloride salts separated at the bottom as an oiland were removed from the funnel. Dichloromethane was removed from themixture at less than 30° C. under vacuum to give an amber oil which wastaken up in hexane (400 mL) and placed in a freezer overnight. Thereaction mixture was then thawed and the soluble portion was filtered toremove the crystals of p-nitrophenyl phosphorodichloridate. The solventwas removed from the filtrate via rotary evaporation at less than 35° C.to give n-hexyl p-nitrophenyl phosphorochloridate.

[0103] A 500 mL flask with stir bar was purged with N₂. The n-hexylp-nitrophenyl phosphorochloridate (20 g, 0.043 mol) in anhydrous THF (25mL) was added, followed by slow addition of a solution of BRIJ® 58(polyoxyethylene(20) cetyl ether; 48.56 g, 0.043 mol) andN-methylimidazole (3.45 mL, 0.043 mol) in anhydrous THF (200 mL). Thereaction mixture was stirred at room temperature for 24 hours. Thesolvent was removed at less than 35° C. by rotary evaporation, and theoily residue was dissolved in methanol (50 mL). A solution ofmethanol/water (85 mL: 15 mL, 200 mL) was added. The solidbis-n,n-dihexyl p-nitrophenyl phosphate was collected by filtration. Themethanol was then stripped off on a rotary evaporator at less than 35°C. Water was removed from the product by lyophilization.

[0104] The Examples in Table 7 can be represented by the followingstructure and were prepared using the above procedure. TABLE 7 BRIJ ®polymers having a terminal p-nitrophenyl phosphate lipase inhibitinggroup with a variety of hydrophobic moieties.

HYDROPHOBIC EXAMPLE POLYMER MOIETY 70 BRIJ ® 98 (n = 19, x = 17)n-dodecyl 71 BRIJ ® 98 (n = 19, x = 17) n-hexadecyl 72 BRIJ ® 35 (n =22, x = 11) n-dodecyl 73 BRIJ ® 35 (n = 22, x = 11) n-hexadecyl 74BRIJ ® 58 (n = 19, x = 15) n-dodecyl 75 BRIJ ® 58 (n = 19, x = 15)n-hexadecyl

EXAMPLE 76

[0105] Preparation of an IGEPAL® polymer having a terminal p-nitrophenylphosphate lipase inhibiting group with a n-hexadecyl hydrophobicmoieties:

[0106] A 500 mL flask with stir bar was purged with N₂, and n-hexadecylp-nitrophenyl phosphorochloridate (20 g, 0.043 mol) in anhydrous THF (25mL) was added, followed by slow addition of a solution of IGEPAL® 720(32.41 g, 0.043 mol) and N-methylimidazole (3.45 mL, 0.043 mol) in THF(200 mL). The reaction mixture was stirred at room temperature for 24hours. The solvent was removed under vacuum at room temperature, andoily product was taken up in methanol (50 mL). A solution ofmethanol/water (85: 15, 200 mL) was added to product. Thebis-n,n-dihexyl p-nitrophenyl phosphate was filtered off, and methanolwas then stripped off under vacuum at less than 35° C. Water was removedfrom the product by lyophilization.

[0107] The Examples in Table 8 can be represented by the followingstructure and were prepared using the above procedure. TABLE 8 IGEPAL ®polymers having a terminal p-nitrophenyl phosphate lipase inhibitinggroup with a variety of hydrophobic moieties.

HYDROPHOBIC EXAMPLE POLYMER MOIETY (R) 76 IGEPAL ® 720 (n = 11)n-dodecyl 77 IGEPAL ® 720 (n = 11) n-hexadecyl 78 IGEPAL ® 890 (n = 39)n-dodecyl 79 IGEPAL ® 890 (n = 39) n-hexadecyl

EXAMPLE 80

[0108] Preparation of [Poly(propylene glycol) block-poly(ethyleneglycol) block-poly(propylene glycol)]polymers having a p-nitrophenylphosphate lipase inhibiting group with a n-hexyl hydrophobic moiety:

[0109] A 500 mL flask with stir bar was purged with N₂, and n-hexylp-nitrophenyl phosphorochloridate (20 g, 0.043 mol) in anhydrous THF (25mL) was added followed by slow addition of a solution of [poly(propyleneglycol) block-poly(ethylene glycol) block-poly(propylene glycol)](average MW=2000, 50 wt. % ethylene glycol; 49.36 g, 0.0215 mol) andN-methylimidazole (3.45 mL, 0.043 mol) in THF (200 mL). The reactionmixture was stirred for 24 hours at room temperature. The solvent wasremoved under vacuum at room temperature, and the oily residue was takenup in methanol (50 mL). A mixture of 85:15 methanol:water solution (200mL) was added, and the bis-n,n-dihexyl p-nitrophenyl phosphateprecipitate was filtered off. Methanol was stripped off by rotaryevaporation at less than 35° C., and water was removed from the productby lyophilization.

[0110] The following Examples were prepared using the above procedure.TABLE 9 Poly(propylene glycol) block-poly(ethylene glycol)block-poly(propylene glycol) polymers (PPG-PEG-PPG) having p-nitrophenylphosphate lipase inhibiting groups with a variety of hydrophobicmoieties. HYDROPHOBIC MOIETY EXAMPLE POLYMER (R) 80 PPG-PEG-PPG 2000hexyl 81 PPG-PEG-PPG 2000 dodecyl 82 PPG-PEG-PPG 2000 hexadecyl

EXAMPLE 83

[0111] Preparation of a PLURONIC® polymer having phosphochloridatelipase inhibiting groups and a decyl hydrophobic moiety:

[0112] After purging with N₂, a solution of phophorousoxychloride (30 g,0.1956 mol) in anhydrous THF (100 mL) was added to a 3 L flask, and themixture was cooled to 0-5° C. A mixture of freshly distilledtriethylamine (27.27 mL, 0.1956 mol) and 1-decanol (30.97 g, 0.1956 mol)in anhydrous THF (300 mL) was added dropwise at a maximum rate of 75mL/hour, keeping the solution temperature at 5° C. After the additionwas complete, a mixture of PLURONIC® (average MW=2900, 142 g, 0.0489mol) and freshly distilled triethylamine (13.7 mL, 0.0978 mol) inanhydrous THF (300 mL) was added at a maximum rate of 75 mL/hour,keeping the solution temperature at 5° C. After the addition wascomplete, the reaction was allowed to warm to room temperature andstirred for 24 hours. The triethylammonium hydrochloride salts wereremoved by filtration. The solvent was removed under vacuum at 30° C.,and the resulting oil was washed with hexane (6×250 mL) to remove theunreacted n-decyl phosphorodichloridate. The product, bis-n-decylphosphorochlorodate PLURONIC®, was dried under high vacuum (0.003 mm Hg)overnight at room temperature.

EXAMPLE 84

[0113] Preparation of a PLURONIC® polymer having N-hydroxysuccinimidylphosphate lipase inhibiting groups and a decyl hydrophobic moiety:

[0114] A 125 mL flask with stir bar was purged with N₂, and a solutionof bis-n-decyl phosphorochlorodate PLURONIC® (prepared as in Example 82;30 g, 0.0178 mol) was added. N-hydroxysuccinimide (2.05 g, 0.0178 mol)was added as a solid and allowed to dissolve. Freshly distilledtriethylamine (2.48 mL, 0.0178 mol) was added, and the reaction mixturewas allowed to stir for 0.5 hours. The triethylammonium hydrochloridesalt was filtered off, and the THF was removed from the filtrate byrotary evaporation at 30° C. The product was dried under high vacuum(0.003 mm Hg) overnight.

EXAMPLE 85

[0115] Preparation of PLURONIC® polymers having pyridoxinyl phosphatelipase inhibiting groups and a decyl hydrophobic moiety:

[0116] A 125 mL flask with stir bar was purged with N₂, and bis-n-decylphosphorochlorodate PLURONIC® (prepared as in Example 82; 30 g, 0.0178mol) in anhydrous dichloromethane (30 mL) was added. Pyridoxinehydrochloride (2.54 g, 0.0178 mol) was added as a solid and allowed todissolve. Freshly distilled triethylamine (4.96 mL, 0.0356 mol) wasadded, and the reaction mixture was allowed to stir for 2 hours. Thetriethylammonium hydrochloride salt was filtered off, and the solventwas removed by a rotary evaporation at less than 35° C. The oil wastaken up in THF (50 mL) and refiltered. The solvent was removed byrotary evaporation, and the product was dried under high vacuum (0.003mm Hg) overnight at room temperature.

[0117] Table 10 lists the polymers prepared in Examples 83, 84 and 85.TABLE 10 PLURONIC ® polymers having a variety of leaving groups.HYDROPHOBIC MOIETY LEAVING GROUP EXAMPLE POLYMER (R) (Y—R¹) 83 PLU 2900decyl chloride 84 PLU 2900 decyl n-hydroxysuccinyl 85 PLU 2900 decylpyridoxinyl

EXAMPLE 86

[0118] Preparation of a PLURONIC® polymer having β-lactone lipaseinhibiting group (Scheme I):

[0119] Intermediate 1:

[0120] 10-Hydroxy methyldecanoate 1 (20 g, 98 mmol), benzyloxy2,2,2-trichloroacetimidate (30 g, 118 mmol), dichloromethane (50 mL) andcyclohexane (100 mL) were added to a 1 L, round-bottomed flask. Themixture was stirred for 5 minutes at room temperature. Trifloromethanesulfonic acid (1.3 mL) was added to the reaction mixture under nitrogenatmosphere. Within a few minutes the temperature rose from roomtemperature to 37° C. The reaction was monitored by TLC (hexane:ethylacetate; 9:1). After 16 hours, the starting material completelydisappeared. The solids were separated from the reaction by filtration,and the filtrate was washed with aqueous saturated sodium bicarbonatesolution (3×100 mL) followed by water (3×100 mL). The organic phase wascollected and dried over anhydrous sodium sulfate. The solvent wasremoved under vacuum at room temperature. The residue was purified onsilica gel column using a gradient of ether/hexane as the mobile phase.The product was eluted from the column in ether-hexane (8:2). Thesolvent was removed in vacuo to yield 10-benzyloxy methyldecanoate(intermediate 1) as a solid (32 g).

[0121] Intermediate 2:

[0122] Intermediate 1 (30 g) was saponified in 6N NaOH solution (100 mL)for 12 hours, then acidified with concentrated HCl. The product wasextracted with chloroform (5×100 mL). The organic layers were combinedand dried over sodium sulfate. The solvent was removed under vacuum togive 10-benzyloxy decanoic acid (intermediate 2) (27 g), which was useddirectly in the next reaction.

[0123] Intermediate 3:

[0124] n-Butyl lithium in hexane (1.6 M solution, 68 mL, 108 mmol) wasadded dropwise to a solution of N,N-diisoproyl amine (15.14 mL, 108mmol) in THF (50 mL) which was maintained at 0° C. After the completionof addition, the mixture was stirred for an additional 10 minutes at 0°C. The mixture was cooled to −50° C., then a solution of intermediate 2(15 g, 54 mmol) in 100 mL of THF was added dropwise. After thecompletion of the addition, the mixture was allowed to warm to roomtemperature, then stirred for 1 hour. The mixture was cooled to −78° C.,and a solution of decyl aldehyde (8.44 g, 54 mmol) in THF (40 mL) wasadded dropwise. After stirring for 3 hours at −78° C., the mixture waswarmed to room temperature, then quenched by addition of saturatedammonium chloride solution (50 mL). The mixture was extracted withdiethyl ether (5×50 mL). The organic layers were combined and dried oversodium sulfate, filtered and evaporated to give intermediate 3 (22 g).

[0125] Intermediate 4:

[0126] Benzenesulfonyl chloride (9.8 g, 56 mmol) was added to a solutionof intermediate 3 (12 g, 28 mmol) in pyridine (200 mL) maintained at 0°C. After addition was complete, the mixture was kept in a refrigeratorat 4° C. for 24 hours, then poured into crushed ice (2 kg) and stirredat room temperature for 20 minutes. The mixture was extracted withdiethyl ether (6×150 mL). The combined organic layers were washed withwater, dried over sodium sulfate, filtered and concentrated in vacuo.The product was purified on a silica gel column using hexane:ethylacetate (9:1) to give intermediate 4 as an oil (9.8 g). IR: 1825⁻¹ cm.

[0127] Intermediate 5:

[0128] Intermediate 4 (9.5 g, 22 mmol) was dissolved in methylenechloride, then hydrogenated under 50 psi of hydrogen for 4 hours using10% Pd/C (1 g) as a catalyst. The solution was filtered, and the solventwas removed under vacuum to give intermediate 5 as an oil (6.9 g).

[0129] Intermediate 6:

[0130] PLURONIC® (MW=1,900; 570 g; 300 mmol) in THF (500 mL) was addeddropwise to a stirred suspension of sodium hydride (15 g) in THF (150mL). After the addition was complete, the mixture was stirred for anadditional 30 minutes at room temperature. A solution of ethyl4-bromobutyrate (117 g, 600 mmol) was added dropwise, and the mixturewas stirred at 60° C. for 16 hours. After cooling to room temperature,the salts were filtered off, and the solvent was removed under vacuum togive light brown viscous material which was suspended in dichloromethane(1 L) and washed with water (3×200 mL). The organic layer was collectedand dried over sodium sulfate, filtered, and the solvent was removedunder vacuum to give intermediate 6 as a viscous liquid (770 g).

[0131] Intermediate 7:

[0132] Intermediate 6 was dissolved in a solution of methanol (1 L) and50% sodium hydroxide solution (100 mL), then stirred for 24 hours atroom temperature. The reaction mixture was acidified with concentratedHCl, and the solvent was removed under vacuum. The residue wasresuspended in dichloromethane (1 L), then washed with water (4×250 mL).The organic layer was dried over sodium sulfate, filtered, and thesolvent was removed under vacuum to give intermediate 7 as a viscousliquid (650 g).

[0133] Intermediate 8:

[0134] 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (4.8g, 25 mmol) was added under nitrogen to a solution of intermediate 7(20.72 g, 10 mmol) in dichloromethane (100 mL) in a round bottom flask.The mixture was stirred for 10 minutes at room temperature, thenN-hydroxy succinimide (2.3 g) was added. The mixture was stirred for 12hours at room temperature, then transferred to a separatory funnel andwas washed with water (3×30 mL). The organic layer was dried overanhydrous sodium sulfate, filtered, and the solvent was removed undervacuum to give 22 g of intermediate 8 which was used directly in thenext step.

EXAMPLE 86

[0135] Triethylamine (3 mL) was added to a solution of intermediate 8(22 g, ˜10 mmol) and intermediate 5 (6.5 g, 20 mmol) in dichloromethane(150 mL). The mixture was stirred for 4 hours at room temperature, thenpoured into a separatory funnel and washed with 5% HCl (3×20 mL) andwater (3×50 mL). The organic layer was dried over sodium sulfate,filtered, and the solvent was removed under vacuum. Example 86 wasobtained as viscous liquid (26 g). This material was used directly inthe in vitro and in vivo assay.

EXAMPLE 87

[0136] Preparation of a PLURONIC® polymer having a disulfide lipaseinhibiting group:

[0137] Intermediate 9:

[0138] 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (1.1 g, 5 mmol) wasadded to a solution of 5,5′-dithiobis(2-nitrobenzoic acid) (3.96 g, 10mmol) in dichloromethane (100 mL). After 10 minutes N-hydroxysuccinimide(0.5 g, 5 mmol) was added, and the reaction was stirred for 6 hours atroom temperature. The reaction mixture was poured into a separatory,then washed with water (3×20 mL). The organic layer was dried overanhydrous sodium sulfate, filtered, and the solvent was removed undervacuum to give intermediate 9 which was used directly in the next step.

[0139] A solution of PLURONIC® (MW=1,900; 9.5 g; 5 mmol) indichloromethane (50 mL), followed by triethylamine (0.5 mL) was added toa solution of intermediate 9 in dichloromethane (100 mL). The mixturewas stirred for 16 hours at room temperature, then poured into aseparatory funnel and washed with water (3×30 mL). The organic layer wasdried over anhydrous sodium sulfate, filtered and The solvent wasremoved under vacuum to give Example 87 as a viscous liquid (12 g).

EXAMPLE 88

[0140] Preparation of a PLURONIC® polymer having an anhydride lipaseinhibiting group:

[0141] Intermediate 10:

[0142] 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (2.2 g, 10 mmol)was added to a solution of 1,2,3-benzene tricarboxylic anhydride (2.1 g,10 mmol) in dichloromethane (100 mL). The mixture was stirred for 10minutes, then N-hydroxysuccinimide (1.0 g, 10 mmol) was added, and thereaction was stirred for 6 hours at room temperature. The reactionmixture was poured into a separatory funnel, then washed with water(3×20 mL). The organic layer was dried over anhydrous sodium sulfate,filtered, and the solvent was removed under vacuum to give intermediate10 which was used directly in the next step.

[0143] A solution of PLURONIC® (MW=1,900; 9.5 g; 5 mmol) andtriethylamine (0.5 mL) in dichloromethane (50 mL) was added to asolution of intermediate 10 in dichloromethane (100 mL). The mixture wasstirred for 16 hours at room temperature, then poured into a separatoryfunnel and washed with water (3×30 mL). The organic layer was dried overanhydrous sodium sulfate, filtered, and the solvent was removed undervacuum to give Example 88 (11.2 g) as viscous liquid.

[0144] In Vitro Assay

[0145] Procedure 1: Tributyrin Substrate

[0146] Potential inhibitors of pancreatic lipase activity were evaluatedusing a titration method employing a pH Stat instrument (RadiometerAmerica, Westlake Ohio). Substrate (1 mL tributyrin) was added to 29.0mL of Tris-HCl buffer (pH 7.0) containing 100 mM NaCl, 5 mM CaCl₂, and 4mM sodium taurodeoxycholate. This solution was stirred for 5 minutesprior to the addition of 210 units of porcine pancreatic lipase (Sigma,21,000 units/mg) dissolved in the assay buffer. The release of butyricacid by the lipase was monitored over a 10 minute period by titratingthe assay system to a constant pH of 7.0 with 0.02 M NaOH. Enzymeactivity was expressed as milliequivalents of base added per minute pergram of enzyme. In subsequent assays, varying amounts of inhibitor weresolubilized in either tributyrin or buffer, depending on the solubilitycharacteristics of the compound, and added to the assay system at timezero.

[0147] Procedure 2: Olive Oil Substrate

[0148] Potential inhibitors of pancreatic lipase activity were evaluatedusing a titration method employing a pH Stat instrument (RadiometerAmerica, Westlake, Ohio). Substrate (15 mL of an olive oil emulsioncontaining 80 mM olive oil and 2 mM oleic acid, dissolved and sonifiedin a buffer consisting of 10 mM Tris-HCl pH 8.0, 110 mM NaCl, 10 mMCaCl₂, 2 mM lecithin, 1.32 mM cholesterol, 1.92 mM sodium glycocholate,1.28 mM sodium taurocholate, 2.88 mM sodium glycodeoxycholate, and 1.92mM sodium taurodeoxycholate) was added to 15 mL of assay buffer(Tris-HCl pH 8.0 containing 110 mM NaCl and 10 mM CaCl₂). This solutionwas stirred for 4 minute prior to the addition of 1050 units of porcinepancreatic lipase (Sigma, 21,000 units/mg) dissolved in assay buffer.The hydrolysis of triglyceride was monitored over a 30 minute period bytitrating the assay system to a constant pH of 8.0 with 0.02M NaOH.Enzyme activity was expressed as milliequivalents of base added perminute per gram of enzyme. In subsequent assays, stock solutions ofinhibitor were prepared in either ethanol or DMSO, and varying amountswere added to the assay system at time zero.

[0149] The assays were conducted as described above using eitherprocedure 1 or 2, and the percent inhibition was derived by comparingthe enzyme activities in the presence and absence of inhibitor. Threeconcentrations of inhibitor were assayed, and the percent inhibition wasplotted against the log of the inhibitor concentration in order todetermine the concentration at which 50% inhibition occurred (IC₅₀). Thefollowing compounds were assayed, with the indicated values for IC₅₀presented in Tables 11-17. TABLE 11 IC₅₀ (μM) IC₅₀ (μM) with with OliveExample Polymer Hydrophobic Moiety Tributyrin Oil Polyethylene glycol(PEG) nitrophenyl phosphates: 10 PEG 1000 pentyl phosphate 400 *** 11PEG 1000 hexyl phosphate na *** 12 PEG 1000 octyl phosphate 538 *** 5PEG 1000 decyl phosphate na *** 6 PEG 1000 dodecyl phosphate 466 *** 7PEG 1000 tetradecyl phosphate 1142 *** 8 PEG 1000 hexadecyl phosphate 67320 9 PEG 1000 octadecyl phosphate 98 *** 13 PEG 1000 docosyl phosphate345 *** 14 PEG 1000 cholesteryl phosphate 172 *** 16 PEG 1500 pentylphosphate na *** 19 PEG 1500 hexadecyl phosphate 215 *** 29 PEG 1500octadecyl phosphate 73 *** 24 PEG 1500 5-phenyl-1-pentyl 24 942phosphate 22 PEG 1500 famesyl phosphate na *** 23 PEG 1500 cholesterylphosphate 307 *** 15 PEG 3400 pentyl phosphate 559 *** 1 PEG 8400 pentylphosphate 455 *** Polypropylene glycol (PPG) nitrophenyl phosphates: 49PPG 1000 pentyl phosphate 4000 *** 53 PPG 2000 pentyl phosphate 52000*** PLURONIC ® polymers having nitrophenyl phosphate: 32 PLU 1100 octylphosphate 61 601 31 PLU 1100 decyl phosphate 174 454 30 PLU 1100 dodecylphosphate 55 400 29 PLU 1100 tetradecyl phosphate 133 1200 33 PLU 1100hexyl phosphate 155 353 39 PLU 1900 pentyl phosphate 3.6 9000 34 PLU1900 octyl phosphate 3.8 379 35 PLU 1900 decyl phosphate 2.4 105 36 PLU1900 dodecyl phosphate 2.3 183 37 PLU 1900 tetradecyl phosphate 3.6 18738 PLU 1900 hexadecyl phosphate 22 196 44 PLU 2900 dodecyl phosphate 1.7286 43 PLU 2900 tetradecyl phosphate 1.7 260 42 PLU 2900 hexadecylphosphate 0.9 106 41 PLU 2900 octadecyl phosphate 1.0 174 48 PLU 4400dodecyl phosphate 8.4 *** 47 PLU 4400 tetradecyl phosphate 5.0 *** 46PLU 4400 hexadecyl phosphate 1.4 *** 45 PLU 4400 octadecyl phosphate 4.8*** 39 PLU 8400 pentyl phosphate 325 *** 40 PLU 8400 hexyl phosphate 84*** Polyethylene glycol (PEG) nitrophenyl phosphonates: 60 PEG 1000pentyl phosphonate 836 na 59 PEG 1500 pentyl phosphonate na ***

[0150] TABLE 12 IC₅₀ values of PLURONIC ® polymers having ap-nitrophenyl phosphate lipase inhibiting group and dialkoxy linkers.HYDROPHOBIC IC₅₀ (mM) IC₅₀ (μM) EXAMPLE PLU MW MOIETY (R) DIALKOXY (Z′)with Tributyrin with Olive Oil 61 1900 n-pentyl n-pent-1,5-dioxy 1.8 na62 1900 n-decyl n-pent-1,5-dioxy 1.1 289 63 1900 n-hexadecyln-pent-1,5-dioxy 1.1 278 66 1900 n-hexadecyl n-undecyl-1,10-dioxy 0.8182

[0151] TABLE 13 IC₅₀ values of polyethylene glycol polymers having ap-nitrophenyl phosphate lipase inhibiting group and dialkoxy linkers.HYDROPHOBIC IC₅₀ (mM) IC₅₀ (μM) EXAMPLE PEG MW MOIETY (R) DIALKOXY (Z′)with Tributyrin with Olive Oil 67 1500 n-hexyl n-pent-1,5-dioxy 71 na 681500 n-dodecyl n-pent-1,5-dioxy 58 371 69 1500 n-hexadecyln-pent-1,5-dioxy 49 184

[0152] TABLE 14 IC₅₀ values for BRIJ ® polymers having a p-nitrophenylphosphate lipase inhibiting group. HYDRO- PHOBIC IC₅₀ (μM) IC₅₀ (μM)MOIETY with with Olive EXAMPLE POLYMER (R) Tributyrin Oil 70 BRIJ ® 98n-dodecyl *** *** (n = 19, x = 17) 71 BRIJ ® 98 n-hexadecyl 250 266 (n =19, x = 17) 72 BRIJ ® 35 n-dodecyl 1800 275 (n = 22, x = 11) 73 BRIJ ®35 n-hexadecyl 1900 392 (n = 22, x = 11) 74 BRIJ ® 58 n-dodecyl 1100 168(n = 19, x = 15) 75 BRIJ ® 58 n-hexadecyl 2200 428 (n = 19, x = 15)

[0153] TABLE 15 IC₅₀ values for IGEPAL ® polymers having a p-nitrophenylphosphate lipase inhibiting group. IC₅₀ (μM) IC₅₀ (μM) EXAMPLE POLYMERHYDROPHOBIC MOIETY (R) with Tributyrin with Olive Oil 76 IGEPAL ® 720 (n= 11) n-dodecyl *** *** 77 IGEPAL ® 720 (n − 11) n-hexadecyl *** *** 78IGEPAL ® 890 (n = 39) n-dodecyl 344 148 79 IGEPAL ® 890 (n = 39)n-hexadecyl *** ***

[0154] TABLE 16 IC₅₀ values for PPG-PEG-PPG polymers havingp-nitrophenyl phosphate lipase inhibiting groups. HYDROPHOBIC IC₅₀ (μM)IC₅₀ (μM) EXAMPLE POLYMER MOIETY (R) with Tributyrin with Olive Oil 81PPG-PEG-PPG 2000 n-dodecyl 2.4 283 82 PPG-PEG-PPG 2000 n-hexadecyl 1.9384

[0155] TABLE 17 IC₅₀ values for PLURONIC ® polymers having n-hexadecylhydrophobes and a variety of leaving groups. EXAMPLE PLU Mol. wt.LEAVING GROUP (Z - R¹) IC₅₀ (μM) with tributyrin IC₅₀ (μM) with OliveOil 83 2900 chloride 0.9 968 84 2900 n-hydroxysuccinyl 0.9 na 85 2900pyridoxinyl 0.09 936

[0156] In Vivo Studies

[0157] Examples 8, 35, 36, 41, 42, 48, 62, 63, 67-69, 71-75, 78, 81 and82 were evaluated for their ability to reduce daily caloric intake byincreasing the excretion of fat in the feces, and to decrease bodyweight gain, relative to the control group, in normal rats over a sixday period. Male Sprague-Dawley rats (five to six weeks of age) wereindividually housed and fed ad libitum a powdered “high fat diet,”consisting of standard rodent chow supplemented with 15% fat (consistingof 55% coconut oil and 45% corn oil) by weight. After feeding theanimals this diet for five days, the animals were weighed and sortedinto the treatment or control groups (6-8 animals per group, each grouphaving equal mean body weights). Animals were treated for six days withthe test compounds, which were added to the “high fat diet” atconcentrations (w/w) of 0.0% (control), 0.3 or 1.0 percent of the diet.

[0158] Food consumption was measured for each animal throughout thestudy, and was expressed as the total amount of food consumed per animalover the six day treatment period. On day 6, each animal was weighed,and the total body weight gain over the treatment period was calculated.

[0159] Rat fecal samples were collected on the final three days of thesix days of drug treatment. The samples were freeze dried and ground toa fine powder. One half gram of sample was weighed and transferred toextraction cells. Samples were extracted in an accelerated solventextractor (ASE 200 Accelerated Solvent Extractor, Dyonex Corporation,Sunnyvale, Calif.) with 95% ethanol, 5% water and 100 mM KOH. The samplewas extracted in 17 minutes at 150° C. and 1500 psi. An aliquot ofextract was transferred to a test tube containing a molar excess of HCl.The sample was then evaporated and reconstituted in a detergent solutionconsisting of 2% Triton X-1200, 1% polyoxyethylene lauryl ether and 0.9%NaCl. Fatty acids were then quantitated enzymatically with acolorimetric kit (NEFAC, Wako Chemical GmbH, Neuss, Germany).

[0160] Table 18 contains values for fecal fat/consumed fat for bothcontrol and test animals (determined enzymatically as described above),and food consumption and body weight gain over 6 days as compared tocontrol animals.

[0161] Calculation of Fecal Fat/Consumed Fat:

[0162] Fatty acid concentrations from the enzymatic assay are expressedas mmol/mL. The mmol/mL of fatty acid is then multiplied by the numberof milliliters of extract generated from 500 mg of sample to give thetotal mmoles of fatty acid. The value for the total mmoles of fatty acidis converted to total milligrams of fatty acid using the averagemolecular weight of medium to long chain fatty acid. The value iscorrected for any dilutions made during sample work-up. When results areexpressed as mgs/gm of feces, the total milligrams of fatty acids ismultiplied by 2. When results are expressed as total milligrams of fattyacid excreted in 24 hours, the mgs/gm of feces value is multiplied byfecal weight in grams excreted in 24 hours. When the results areexpressed as excreted fat as a percentage of that consumed in 24 hours,the total weight of fat excreted in 24 hours is divided by the weight offatty acids consumed in over 24 hours and multiplied by 100. TABLE 18 Invivo results of selected polymers having lipase inhibiting groups. Totalfood Total weight Compound Fecal fat consumption change Number ClassBackbone Hydrophobe % of consumed % of control % of control 8 phosphatePEG 1000 hexadecyl   2 ± 0.5    87 ± 2.8**   66 ± 9.8** 67 phosphate C5PEG 1500 hexyl   3 ± 0.7   97 ± 7.8  127 ± 64.4 68 phosphate C5 PEG 1500dodecyl   2 ± 0.5   99 ± 12.2   82 ± 15.4* 69 phosphate C5 PEG 1500hexadecyl   3 ± 0.8  105 ± 5.5  92 ± 8.7 35 phosphate PLU 1900 decyl 23± 5    58 ± 10**  −9 ± 17** 36 phosphate PLU 1900 dodecyl 12 ± 3   62 ±7**  16 ± 18** 42 phosphate PLU 2900 hexadecyl   13 ± 2.5**    86 ±8.6**    75 ± 16.1** 41 phosphate PLU 2900 octadecyl   15 ± 3.5**   91 ±6.2*   82 ± 6.6** 48 phosphate PLU 4400 dodecyl  4 ± 1** 96 ± 7  79 ±8**  81 phosphate PPG-PEG-PPG dodecyl 2 ± 0 92 ± 8  78 ± 11** 82phosphate PPG-PEG-PPG hexadecyl 2 ± 0 114 ± 8   129 ± 12** 62 phosphateC5 PLU 1900 decyl  12 ± 3.2   47 ± 2.6   −24 ± 13.6** 63 phosphate C5PLU 1900 hexadecyl  6 ± 1**  90 ± 5**    77 ± 13.3** 71 phosphate Brij98 hexadecyl 2 ± 1 98 ± 6 85 ± 11 72 phosphate Brij 35 dodecyl 1 ± 0 95± 5  73 ± 13** 73 phosphate Brij 35 hexadecyl 1 ± 0 106 ± 10  122 ± 17**74 phosphate Brij 58 dodecyl 2 ± 0  90 ± 5** 61 ± 14 75 phosphate Brij58 hexadecyl 1 ± 0 104 ± 8  100 ± 12  78 phosphate Igepal 890 dodecyl 1± 0 93 ± 5  72 ± 13** Control 2 − 3 100 100

[0163] While this invention has been particularly shown and describedwith references to preferred embodiments thereof, it will be understoodby those skilled in the art that various changes in form and details maybe made therein without departing from the spirit and scope of theinvention as defined by the appended claims.

What is claimed is:
 1. A method for treating obesity in a mammal,comprising the step of orally administering to the mammal an effectiveamount of a polymer substituted with at least one group having thefollowing structure:

wherein, R is a hydrogen, hydrophobic moiety, —NR²R³, —CO₂H, —OCOR²,—NHCOR², a substituted or unsubstituted aliphatic group or a substitutedor unsubstituted aromatic group; Z is an oxygen, alkylene, sulfur,—SO₃—, —CO₂—, —NR²—, —CONR²—, —PO₄H— or a spacer group; R² and R³ are,independently, a hydrogen, a substituted or unsubstituted aliphaticgroup, or a substituted or unsubstituted aromatic group; and p is aninteger from zero to about
 30. 2. A method for treating obesity in amammal, comprising the step of orally administering to the mammal aneffective amount of a polymer substituted with at least one group havingthe following structure:

wherein, ring A is a substituted or unsubstituted cyclic aliphatic groupor an aromatic group, or a combination thereof, having one or moreheteroatoms; Z is an oxygen, alkylene, sulfur, —SO₃—, —CO₂—, —NR²—,—CONR²—, —PO₄H— or a spacer group; R² is a hydrogen, a substituted orunsubstituted aliphatic group, or a substituted or unsubstitutedaromatic group; and X is, independently, —PO₂—, —SO₂— or —CO—.
 3. Amethod for treating obesity in a mammal, comprising the step of orallyadministering to the mammal an effective amount of a polymer substitutedwith at least one group having the following structure:

wherein, R is a hydrogen, hydrophobic moiety, —NR²R³, —CO₂H, —OCOR²,—NHCOR², a substituted or unsubstituted aliphatic group or a substitutedor unsubstituted aromatic group; Z is an oxygen, alkylene, sulfur,—SO₃—, —CO₂—, —NR²—, —CONR²—, —PO₄H— or a spacer group; R² and R³ are,independently, a hydrogen, a substituted or unsubstituted aliphaticgroup, or a substituted or unsubstituted aromatic group; X is,independently, —PO₂—, —SO₂— or —CO—; k is 0 to about 10; and p is aninteger from zero to about
 30. 4. A method for treating obesity in amammal, comprising the step of orally administering to the mammal aneffective amount of a polymer substituted with at least one group havingthe following structure:

wherein, —CO₂H and —OH substituents on the phenyl ring are ortho or parato each other; R⁵ is a hydrophobic moiety, a substituted orunsubstituted aliphatic group or a substituted or unsubstituted aromaticgroup; Z is an oxygen, alkylene, sulfur, —SO₃—, —CO₂—, —NR²—, —CONR²—,—PO₄H— or a spacer group; and R² is a hydrogen or an unsubstituted orsubstituted aliphatic group, or unsubstituted aryl or substitutedaromatic group.
 5. A method for treating obesity in a mammal, comprisingthe step of orally administering to the mammal an effective amount of apolymer substituted with at least one group having the followingstructure:

wherein, R is a hydrogen, hydrophobic moiety, —NR²R³, —CO₂H, —OCOR²,—NHCOR², a substituted or unsubstituted aliphatic group or a substitutedor unsubstituted aromatic group; W¹ and W² are each independently ahalogen or a hydrogen wherein at least one of W¹ or W² is a halogen; Yis oxygen, sulfur, NR² or is absent; and R² and R³ are, independently, ahydrogen, an unsubstituted or substituted aliphatic group, orunsubstituted or substituted aromatic group.
 6. A method for treatingobesity in a mammal, comprising the step of orally administering to themammal an effective amount of a polymer substituted with at least onegroup having the following structure:

wherein, R is a hydrogen, hydrophobic moiety, —NR²R³, —CO₂H, —OCOR²,—NHCOR², a substituted or unsubstituted aliphatic group or a substitutedor unsubstituted aromatic group; Z is an oxygen, alkylene, sulfur,—SO₃—, —CO₂—, —NR²—, —CONR²—, —PO₄H— or a spacer group; R² and R³ are,independently, a hydrogen or an unsubstituted or substituted aliphaticgroup, or an unsubstituted or substituted aromatic group; and p is aninteger from zero to about
 30. 7. The method of claim 6, wherein thepolymer is a fat-binding polymer.